efficient-agroinfiltration-plants-for-high-level-transient-expression

Efficient Agroinfiltration of Plants for High-level Transient Expression of Recombinant Proteins

Mammalian cell culture is the major platform for commercial production of human vaccines and therapeutic proteins. However, it cannot meet the increasing ...

Arizona State University 

While toxicokinetics has become an integral part of pharmaceutical safety assessment over the last two decades, its use in the chemical industry is relatively new. However, it is recognised as a potentially important tool in human health risk assessment and recent initiatives have advocated greater application of toxicokinetics as part of an improved assessment strategy for crop protection chemicals that could offer greater efficiency, use fewer animals and provide better data for risk assessment purposes. To explore the potential scientific and animal welfare benefits of increased use of toxicokinetic data across the chemical industry, an international workshop was held in 2008. Experts from a wide range of chemical industry sectors, including industrial chemicals, agrochemicals and consumer products, participated in the meeting as well as representatives from relevant regulatory authorities. Pharmaceutical industry experts were also invited, in order to share experiences from the extensive use of toxicokinetics in drug development. Given that increased generation of toxicokinetic data could potentially result in an increased number of animals undergoing testing, technologies and strategies to reduce and refine animal use for this purpose were also considered. This paper outlines and expands upon the key themes that emerged from the workshop.

Application of toxicokinetics to improve chemical risk assessment: implications for the use of animals.

Alors que la toxicocin&eacute;tique est devenu partie int&eacute;grante de l&#39;&eacute;valuation de la s&eacute;curit&eacute; pharmaceutique au cours des deux derni&egrave;res d&eacute;cennies, son utilisation dans l&#39;industrie chimique est relativement nouvelle. Cependant, elle est reconnue comme un outil potentiellement important dans l&#39;&eacute;valuation des risques sanitaires et des initiatives r&eacute;centes ont pr&eacute;conis&eacute; une plus grande demande de la toxicocin&eacute;tique dans le cadre d&#39;une strat&eacute;gie d&#39;&eacute;valuation am&eacute;lior&eacute;e pour produits chimiques protection des cultures qui pourrait offrir une plus grande efficacit&eacute;, utiliser moins d&#39;animaux et fournir de meilleures donn&eacute;es aux fins de l&#39;&eacute;valuation des risques. Pour explorer les b&eacute;n&eacute;fices potentiels de scientifiques et de bien-&ecirc;tre animal de l&#39;utilisation accrue des donn&eacute;es toxicocin&eacute;tiques dans l&#39;ensemble de l&#39;industrie chimique, un atelier international a eu lieu en 2008. Experts d&#39;un large &eacute;ventail de secteurs de l&#39;industrie chimique, y compris les produits chimiques industriels, de produits agrochimiques et de produits de consommation, ont particip&eacute; &agrave; la r&eacute;union ainsi que des repr&eacute;sentants des autorit&eacute;s r&eacute;glementaires comp&eacute;tentes. Experts de l&#39;industrie pharmaceutique ont &eacute;galement &eacute;t&eacute; invit&eacute;s, afin de partager les exp&eacute;riences de l&#39;utilisation &eacute;tendue de la toxicocin&eacute;tique, d&eacute;veloppement des m&eacute;dicaments. Compte tenu de cette production accrue de donn&eacute;es pourraient entra&icirc;ner un augmentation du nombre d&#39;animaux qui subissent des tests de toxicocin&eacute;tique, technologies et strat&eacute;gies de r&eacute;duction et d&#39;affiner des animaux utilis&eacute;s &agrave; cette fin ont &eacute;galement &eacute;t&eacute; examin&eacute;s. Ce document pr&eacute;sente et d&eacute;veloppe les th&egrave;mes cl&eacute;s qui ont &eacute;merg&eacute; de l&#39;atelier.

Application de la toxicocin&eacute;tique d&#39;am&eacute;liorer l&#39;&eacute;valuation des risques chimiques : implications pour l&#39;utilisation des animaux.

Mentre tossicocinetica &egrave; diventata parte integrante della valutazione di sicurezza farmaceutica negli ultimi due decenni, il suo utilizzo nell&#39;industria chimica &egrave; relativamente nuovo. Tuttavia, &egrave; riconosciuta come uno strumento potenzialmente importante nella valutazione del rischio salute umana e recenti iniziative hanno auspicato una maggiore applicazione della tossicocinetica come parte di una strategia di valutazione migliorata per le sostanze chimiche di protezione delle colture che potrebbe offrire una maggiore efficienza, utilizzare un minor numero di animali e forniscono dati migliori per scopi di valutazione del rischio. Per esplorare i potenziali benefici del benessere animale e scientifico dell&#39;aumento dell&#39;uso di dati tossicocinetici in tutta l&#39;industria chimica, nel 2008 si &egrave; tenuto un workshop internazionale. Esperti provenienti da una vasta gamma di settori dell&#39;industria chimica, compresi i prodotti chimici industriali, prodotti chimici e prodotti di consumo, ha partecipato a riunioni, nonch&eacute; di rappresentanti dalle competenti autorit&agrave; di regolamentazione. Sono stati anche invitati esperti del settore farmaceutico, al fine di condividere esperienze dall&#39;uso estensivo della tossicocinetica in drug development. Dato che maggiore generazione di tossicocinetici dati potenzialmente potrebbero tradursi in un aumento del numero di animali sottoposti a test, tecnologie e strategie per ridurre e raffinare l&#39;uso di animali per questo scopo sono stati anche considerate. Questo documento delinea e amplia i temi chiave emersi dal workshop.

Applicazione della tossicocinetica per migliorare la valutazione del rischio chimico: implicazioni per l&#39;uso di animali.

Toxicokinetic (TK) information can substantially enhance the value of the data generated from toxicity testing, and is an integral part of pharmaceutical safety assessment. It is less widely used in the chemical, agrochemical and consumer products industries, but recognition of its value is growing, as reflected by increased reference to the use of TK information in new and draft OECD test guidelines. To help promote increased consideration of the important role TK can play in chemical risk assessment, we have gathered practical examples from the peer-reviewed literature, as well as in-house industry data, that highlight opportunities for the use of TK in the selection of dose levels. Use of TK can help to ensure studies are designed to be of most relevance to assessing potential risk in humans, and avoid the use of excessively high doses that could result in unnecessary suffering in experimental animals. Greater emphasis on the potential contribution of TK in guiding study design and interpretation should be incorporated in regulatory data requirements and associated guidance.

Use of toxicokinetics to support chemical evaluation: Informing high dose selection and study interpretation.

Informations toxicocin&eacute;tiques (TK) peuvent augmenter consid&eacute;rablement la valeur des donn&eacute;es produites des tests de toxicit&eacute; et fait partie int&eacute;grante de l&#39;&eacute;valuation de la s&eacute;curit&eacute; pharmaceutique. Il est moins couramment dans les industries chimiques, agrochimiques et consommateur de produits, mais la reconnaissance de sa valeur augmente, comme en t&eacute;moigne une augmentation r&eacute;f&eacute;rence &agrave; l&#39;utilisation de savoirs traditionnels information &agrave; nouveau et le projets OECD test guidelines. Pour aider &agrave; promouvoir un examen accru le r&ocirc;le important de que savoirs traditionnels peuvent jouer dans l&#39;&eacute;valuation des risques chimiques, nous avons rassembl&eacute; des exemples pratiques depuis les pairs, ainsi que les donn&eacute;es internes de l&#39;industrie, qui mettent en &eacute;vidence les possibilit&eacute;s d&#39;utilisation des savoirs traditionnels dans le choix des niveaux de dose. Utilisation des savoirs traditionnels peut aider &agrave; s&#39;assurer que les &eacute;tudes sont con&ccedil;ues pour &ecirc;tre des &eacute;l&eacute;ments les plus pertinents pour &eacute;valuer le risque potentiel chez l&#39;homme et &eacute;vitez d&#39;utiliser des doses excessivement &eacute;lev&eacute;es qui pourraient entra&icirc;ner des souffrances chez les animaux exp&eacute;rimentaux inutiles. Davantage l&#39;accent sur la contribution potentielle des savoirs traditionnels dans le plan directeur de l&#39;&eacute;tude et l&#39;interpr&eacute;tation devrait &ecirc;tre incluse dans les donn&eacute;es r&eacute;glementaires exig&eacute;es et associ&eacute;s d&#39;orientation.

Utilisation de la toxicocin&eacute;tique d&#39;appuyer l&#39;&eacute;valuation chimique : informer l&#39;interpr&eacute;tation de s&eacute;lection et &eacute;tude de dose &eacute;lev&eacute;e.

Informazioni tossicocinetici (TK) possono migliorare sostanzialmente il valore dei dati generati dai test di tossicit&agrave; ed sono parte integrante della valutazione di sicurezza farmaceutica. &Egrave; meno ampiamente usato nelle industrie di prodotti chimici, agrochimico e dei consumatori, ma riconoscimento del suo valore &egrave; in crescita, come riflesso di maggiore riferimento all&#39;uso di informazioni di TK in bozza e nuove test linee guida dell&#39;OCSE. Per aiutare a promuovere maggiore considerazione del ruolo importante che TK pu&ograve; giocare nella valutazione del rischio chimico, abbiamo raccolto esempi pratici di letteratura peer-reviewed, cos&igrave; come i dati interni dell&#39;industria, che mettono in risalto le opportunit&agrave; per l&#39;utilizzo di TK nella selezione dei livelli di dose. Uso di TK pu&ograve; contribuire a garantire che gli studi sono progettati per essere pi&ugrave; rilevante per la valutazione del potenziale rischio negli esseri umani ed evitare l&#39;uso di dosi troppo elevate che potrebbero provocare inutili sofferenze in animali da esperimento. Maggiore enfasi sul potenziale contributo di TK in guida design studio e interpretazione dovrebbe essere incorporata nei requisiti dei dati normativi e associati di orientamento.

Uso della tossicocinetica per supportare la valutazione chimica: informare l&#39;interpretazione di selezione e studio di dose elevata.

As part of a longer-term goal to create a quantitative mechanistic model of the p53-Mdm2 DNA-damage pathway, we are studying cellular responses to compounds causing DNA-damage by various modes-of action, including two natural polyphenols: quercetin (QUE) and curcumin (CUR). QUE and CUR are weak mutagens in some in vitro assays and possess both anti- or pro-oxidant effects depending on dose. This study examines the dose-response of DNA-damage pathway to these compounds in HT1080 cells (a human cell line with wild-type p53) at doses relevant to human exposure. CUR was more potent in causing reactive oxygen species, DNA damage (measured as phospho-H2AX) and p53 induction, with lowest observed effect levels (LOELs; 3-8Î¼M) approximately three-fold lower than QUE (20-30Î¼M). CUR showed a strong G2/M arrest and apoptosis at âˆ¼10Î¼M. QUE caused S phase arrest at low doses (8Î¼M) and apoptosis was only induced at much higher doses (60Î¼M). At concentrations with similar levels of p-H2AX and p53 biomarkers, CUR caused greater micronuclei frequency. CUR induced clear increases micronuclei at 3-6Î¼M, while QUE had a weaker micronuclei response even at the highest doses. Thus, even with two compounds sharing common chemistries, DNA-damage response patterns differed significantly in terms of dose and cell fate.

Assessing dose-dependent differences in DNA-damage, p53 response and genotoxicity for quercetin and curcumin.

Dans le cadre d&#39;un objectif &agrave; plus long terme pour cr&eacute;er un mod&egrave;le quantitatif m&eacute;caniste de la voie de dommages &agrave; l&#39;ADN Mdm2-p53, nous &eacute;tudions les r&eacute;ponses cellulaires aux compos&eacute;s &agrave; l&#39;origine des l&eacute;sions de l&#39;ADN par divers modes-d&#39;action, y compris deux polyph&eacute;nols naturels : querc&eacute;tine (Qu&eacute;.) et la curcumine (CUR). QUE et CUR sont mutag&egrave;nes faibles dans certains essais in vitro et poss&egrave;dent les deux effets anti - ou pro - antioxydants selon la dose. Cette &eacute;tude examine la relation dose-effet de voie dommages &agrave; l&#39;ADN de ces compos&eacute;s dans les cellules HT1080 (une lign&eacute;e cellulaire humaine avec p53 sauvage) &agrave; des doses pertinentes &agrave; l&#39;exposition humaine. Actu &eacute;tait plus puissant dans l&#39;apparition des esp&egrave;ces r&eacute;actives de l&#39;oxyg&egrave;ne, l&eacute;sions de l&#39;ADN (mesur&eacute;e en phospho-H2AX) et l&#39;induction de p53, avec les niveaux de l&#39;effet observ&eacute; (DMEO; 3-8&mu;M) environ moins tripl&eacute; que Qu&eacute;bec (20-30&mu;M). CUR a montr&eacute; une forte G2/M arrestation et l&#39;apoptosis &agrave; &sim;10&mu;M. QUE fait arrestation phase S &agrave; faible dose (8&mu;M) et l&#39;apoptose a &eacute;t&eacute; induite uniquement &agrave; des doses beaucoup plus &eacute;lev&eacute;es (60&mu;M). &Agrave; des concentrations &agrave; des niveaux similaires de biomarqueurs p-H2AX et p53, CUR a caus&eacute; une plus grande fr&eacute;quence de micronoyaux. Actu induit clairement augmente micronoyaux &agrave; 3-6&mu;M, tandis que le Qu&eacute;bec avait une r&eacute;ponse plus faible des micronoyaux m&ecirc;me aux doses maximales. Ainsi, m&ecirc;me avec deux compos&eacute;s partage communs chimies, profils de r&eacute;ponse des l&eacute;sions de l&#39;ADN diff&egrave;rent significativement en fonction de la dose et cell fate.

D&eacute;terminer les diff&eacute;rences de dose-d&eacute;pendante dans les l&eacute;sions de l&#39;ADN, de r&eacute;ponse p53 et de g&eacute;notoxicit&eacute; de querc&eacute;tine et de la curcumine.

Article	Year
Application of toxicokinetics to improve chemical risk assessment: implications for the use of animals. Regulatory toxicology and pharmacology : RTP\| PubMed ID: 19665509	2009
Use of toxicokinetics to support chemical evaluation: Informing high dose selection and study interpretation. Regulatory toxicology and pharmacology : RTP\| PubMed ID: 22198561	2012
Assessing dose-dependent differences in DNA-damage, p53 response and genotoxicity for quercetin and curcumin. Toxicology in vitro : an international journal published in association with BIBRA\| PubMed ID: 23764886	2013

Matthew Dent

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